Processing hundreds of SARS-CoV-2 samples with an in-house PCR-based method without robotics
Sprache des Titels:
Englisch
Original Kurzfassung:
The SARS-CoV-2 pandemic has required the development of multiple testing systems to moni-tor and control the viral infection. Here, we developed a PCR test to screen COVID-19 infections that can process up to ~180 samples per day without the requirement of robotics. For this pur-pose, we implemented the use of multi-channel pipettes and plate magnetics for the RNA extrac-tion step and combined the reverse transcription step with the qPCR within one step. We tested the performance of two RT-qPCR kits, as well as different sampling buffers and showed that samples taken in NaCl or PBS are stable and compatible with different COVID-19 testing sys-tems. Finally, we designed a new internal control based on the human RNase P gene that does not require a DNA digestion step. Our protocol is easy to handle and reaches the sensitivity and accuracy of the standardized diagnostic protocols used in the clinic to detect COVID-19 infec-tions.
Forschungs-